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Zhang et al. Gut Pathog (2017) 9:11 DOI
10.1186/s13099-017-0160-6
RESEARCH
Oral administration of Clostridium butyricum CGMCC0313-1
inhibits β-lactoglobulin-induced intestinal anaphylaxis in a
mouse model of food allergyJuan Zhang1, Hui Su2, Qiuhong Li1,
Haixia Wu1, Mengyun Liu3, Jianqiong Huang3, Minghua Zeng1, Yuejie
Zheng3* and Xin Sun1*
Abstract Background: Probiotic bacteria can induce immune
regulation or immune tolerance in patients with allergic diseases,
but the underlying mechanisms are still unclear. There has been a
growing interest in the use of beneficial bacteria for allergic
diseases recently. This study aimed at exploring whether
Clostridium butyricum CGMCC0313-1 (C. butyricum) can reduce
β-lactoglobulin(BLG)-induced intestinal anaphylaxis in a murine
model of food allergy.
Methods: The preventive and therapeutic effects of oral C.
butyricum on anaphylactic symptoms induced via BLG in food allergy
mice were investigated. Intestinal anaphylaxis, T helper
(Th)-specific cytokines and transcription factors, secretory IgA
(sIgA), CD4+ CD25+ Foxp3Treg cell and histopathological alterations
were examined.Results: Clostridium butyricum significantly
ameliorated intestinal anaphylaxis symptoms in the food allergy
mice. sIgA and CD4+ CD25+ Foxp3Treg cell were increased by oral C.
butyricum. It also reversed the imbalance of Th1/Th2
andTh17/Treg.
Conclusions: Clostridium butyricum reduces BLG-induced
intestinal anaphylaxis in mice and might be an additional or
supplementary therapy for food allergy.
Keywords: β-Lactoglobulin, Clostridium butyricum, Food allergy,
Mice, Probiotics
© The Author(s) 2017. This article is distributed under the
terms of the Creative Commons Attribution 4.0 International License
(http://creativecommons.org/licenses/by/4.0/), which permits
unrestricted use, distribution, and reproduction in any medium,
provided you give appropriate credit to the original author(s) and
the source, provide a link to the Creative Commons license, and
indicate if changes were made. The Creative Commons Public Domain
Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the
data made available in this article, unless otherwise stated.
BackgroundFood is a foreign antigen that is necessary for
nutrition. Inevitably, food antigens present a continuous challenge
throughout life. Humans have adapted to food via mech-anisms of
immune tolerance. Food allergy is defined as abnormal immune
responses resulting from breakdown of natural oral tolerance [1].
Food allergy is an increas-ing public health problem worldwide
[2–5] and has been estimated to affect approximately 5% of adults
and 8% of children [6]. Among food allergies, cow’s milk allergy
is
one of the earliest and most prevalent food allergies and
β-lactoglobulin (BLG) is the major allergen [7, 8]. Diets based on
cow’s milk play a major role in children’s nutri-tion as it has an
essential effect on the patient’s quality of life. Additionally, up
to now the current standard for pre-vention of food allergy is
still the strict allergen avoidance and the elimination of the
triggering food from the diets [9]. However,
accidental ingestion is difficult to be abso-lutely avoided in
our life. Therefore, effectively prevent and manage food allergy to
restore immune tolerance is particularly needed.
A previous study shows that the increased prevalence of allergic
diseases is associated with decreased micro-bial exposure and
alteration of microbial communities represented in the gut
microbiota [10]. It has been dem-onstrated that the composition and
metabolic activity of
Open Access
Gut Pathogens
*Correspondence: [email protected]; [email protected] 1
Department of Pediatrics, Xijing Hospital, the Fourth Military
Medical University, Xi’an 710032, China 3 Respiratory Department,
Shenzhen Children’s Hospital, Shenzhen 518036, ChinaFull list of
author information is available at the end of the article
http://creativecommons.org/licenses/by/4.0/http://creativecommons.org/publicdomain/zero/1.0/http://creativecommons.org/publicdomain/zero/1.0/http://crossmark.crossref.org/dialog/?doi=10.1186/s13099-017-0160-6&domain=pdf
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Page 2 of 10Zhang et al. Gut Pathog (2017) 9:11
the microbiota is crucial for both the maintenance and the
development of immune homeostasis, as well as the induction of
immune tolerance [11, 12]. Intervention strategies targeting the
intestinal microbiota include the deliberate administration of
probiotic bacteria. Probiot-ics are live microorganisms that confer
a health benefit to the host when administered in adequate amounts
[13]. Different bacterial strains or their mixtures have been used
to assess their protective effects for allergic diseases in
clinical trials, but the results have been controversial [14–16].
Possible mechanisms of their protective action include both the
induction of regulatory dendritic cells (regDCs) and T cells and
the skewing the imbalances of T helper (Th)1/Th2 as well as
Th17/Treg, together with the enhancement of the epithelial barrier
func-tion [10, 17–21]. Imbalances in Th responses can also be
detected using Th-specific transcription factors: T-bet for Th1
cells, GATA-3 for Th2 cells, retinoic acid orphan receptor-γt
(RORγt) for Th17 cells and forkhead box P3 (Foxp3) for Tregs [22].
Nevertheless, the knowledge of molecular mechanisms underlying
probiotic-host inter-actions that shape host immune system in a
protective setting is still incomplete. Mouse models of food
allergy to clinically relevant allergens could be helpful in
provid-ing information difficult to be obtained in human
studies.
Clostridium butyricum CGMCC0313-1 (C. butyricum) has been widely
used for improving gastrointestinal func-tion as probiotics [23,
24]. However, the benefit of C. butyricum on food allergy is rarely
reported. This study aimed to explore whether oral C. butyricum can
reduce food allergy in mice. Findings from this study will
con-tribute to a better understanding of the protective effects of
the beneficial bacteria in allergic diseases.
MethodsMiceMale BALB/c mice of 6–8 weeks were purchased
from the Laboratory Animal Center of the Fourth Mili-tary Medical
University and acclimated to their new environment for 1
week. Animals were housed under conventional conditions, fed
standard mouse pellets and water adlibitum. Experimental
procedures were approved by the Ethics Committee for Animal Studies
of the Fourth Military Medical University (20150901) and performed
in accordance with their guidelines of the Institutional Animal
Care and Use Committee.
The probioticsThe C. butyricum powder (Kexing Biotech Company
lim-ited, Weifang, Shandong, China) was stored at −20 °C.
Drinks were prepared using normal saline (NS) only or NS plus C.
butyricum. The concentration of C. butyricum was
5 × 108 CFU/ml.
Mouse model of food allergyBALB/c mice were randomly
assigned into 4 experimen-tal groups with 10 in each group. The 4
groups of mice were treated as follow: the food allergy group
received 20 mg BLG (Sigma, St. Louis, MO, USA) plus 10 μg
chol-era toxin(CTX, Sigma) orally on days 7, 14 and 21, and
followed by oral administration of 100 mg BLG chal-lenge on
day 28; the preventive and treated groups were approached as the
food allergy group, and the animals were given 200 μl C.
butyricum feeding from days 1 to 21 or from days 22 to 28
respectively; mice from the control group only received 10 μg
CTX orally, and followed by challenge with NS (Fig. 1).
Sample collection
Challenge: BLG or
NS
Sensitization: BLG
plus CTX or CTX
C. butyricum C. butyricum
D0 D7 D14 D21 D28 D29
Prevention: Treatment:
Fig. 1 The protocols used for the mouse models of food allergy.
Male BALB/c mice were sensitized orally with BLG plus CTX or only
CTX on day 7, 14 and 21, and followed by oral administration of 100
mg BLG challenge on day 28. The animals received C. butyricum
feeding from day 1 to day 21 or from day 22 to day 28 by oral
gavage
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Measurement of intestinal anaphylaxisIntestinal anaphylaxis
was assessed in challenged mice by the allergic diarrhea and
circulating mouse mast cell protease 1 (mMCP-1) levels as
previously described [25, 26]. Briefly, mice were observed for the
presence of diar-rhea for 1 h after the challenge and were
scored as diar-rhea positive or negative. Diarrhea and anaphylactic
symptoms were scored by visually monitoring mice for 60 min
after challenge. Diarrhea—0, normal stools; 1, a few wet and
unformed stools; 2, a number of wet and unformed stools with
moderate perianal staining of the coat; 3, severe, watery stool
with severe perianal staining of the coat. Anaphylactic symptoms—0,
no symptoms; 1, reduced activity, trembling of limbs; 2, loss of
con-sciousness, no activity upon prodding; 3, convulsions, death.
Serum was obtained 1 h following the final antigen challenge
for measurement of mMCP-1 using the com-mercial enzyme-linked
immunosorbentassay (ELISA) (eBioscience, San Diego, CA, USA)
according to manu-facturer instructions (Table 1).
The measurement of cytokines and immunoglobulinSerum
samples were collected to assay the presence of cytokines and
immunoglobulin (Ig). Levels of serum IL-4, IL-5, IL-13, IL-17,
INF-γ, IL-10, TGF-β1 and the total IgE were measured by ELISA Kits
(RD system, Boston, MA, USA; Uscn Life, Wuhan, Hubei, China)
following the manufacture’s protocol.
Secretory IgASmall intestine was rinsed with 10 ml of cold
PBS. Intes-tinal lavages were centrifuged at 12,000×g for
20 min at 4 °C, and levels of secretory IgA (sIgA) in
the superna-tants were determined by ELISA (Uscn Life) as
previ-ously described [27].
RNA isolation and quantitative real‑time PCRAfter mice were
sacrificed on day 29, the spleen was dis-sected and immersed in
RNAlater® (Ambion, Austin, TX, USA) for PCR. Total RNA
(n = 6 mice per group) was carried out using Trizol
(Ambion) from whole spleen tis-sue and reverse-transcribed to cDNA
by a PrimeScript™ RT Master Mix Kit (TaKaRa, Tokyo, Japan). cDNA
was amplified using SYBR® Premix Ex Taq™ II Kit (TaKaRa)
and run in the Real-Time PCR Detection System. Primers for T
cell transcription factors were designed and syn-thesized by
TaKaRa Biotechnology (Dalian, Liaoning, China). The sequences were
listed in Table 2. The relative levels of gene expression were
calculated by reference to the level of β-actin using the ΔΔCT
method [28].
Flow cytometrySingle-cell suspensions isolated from mesenteric
lymph nodes (MLN) were stained for FACS analyses as described
previously [29]. Cells were first stained for sur-face markers
including CD4-PerCP-Cy5.5, CD25-FITC (BD Pharmingen, San Diego, CA,
USA). If required, cells were then fixed and permeabilized by BD
Cytofix/Cytop-erm reagent (BD Bioscience, San Jose, CA, USA) and
stained for intracellular expression markers, Foxp3-PE. Data were
acquired with FACSCanto (Beckman Coulter, Miami, FL, USA) and
analyzed by FlowJo 10.0.7 software.
Histological analysisParaffin-embedded sections of proximal
jejunum were stained with hematoxylin and eosin (H&E) for
morpho-logical analysis. Villus length was determined by meas-uring
the distance in μm from the crypt neck to the villus tip using
Image J software. Six animals from each experimental group were
evaluated, and a minimum of 12 well-oriented villi from each
section were measured. Data were reported as villus size measured
in μm. Mor-phological analyses were performed in a blinded manner
to prevent observer bias.
Table 1 Scoring methods of diarrhea and anaphylactic
symptoms
Diarrhea Anaphylactic symptoms Scores
Normal stools No symptoms 0
A few wet and unformed stools Reduced activity, trembling of
limbs 1
A number of wet and unformed stools with moderate perianal
staining of the coat Loss of consciousness, no activityupon
prodding 2
Severe, watery stool with severe perianal staining of the coat
Convulsions, death 3
Table 2 Oligonucleotide primers used in the study
F forward, R reverse
T-bet(F) 5′-CATGGAGAACGGAGAATGGA-3′
T-bet(R) 5′-TGGACAGGGGAAGAGAGCA-3′
RORγt(F) 5′-GCTCCATATTTGACTTTTCCCACT-3′
RORγt(R) 5′-GATGTTCCACTCTCCTCTTCTCTTG-3′
GATA-3(F) 5′-GGATTTAAGTCGAGGCCCAAG-3′
GATA-3(R) 5′-ATTGCAAAGGTAGTGCCCGGTA-3′
Foxp3(F) 5′-CCCAGGAAAGACAGCAACCTT-3′
Foxp3(R) 5′-TTCTCACAACCAGGCCACTTG-3′
β-actin(F) 5′-CATCCGTAAAGACCTCTATGCCAAC-3′
β-actin(R) 5′-ATGGAGCCACCGATCCACA -3′
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Statistical analysisData were expressed as the
mean ± standard error means (SEMs). All data were
analyzed with SPSS17.0 software (SPSS Inc, Illinois, Chicago, USA).
One-Way ANOVA and Mann–Whitney U non-parametric test was con-ducted
to determine the statistical significance, where appropriate. A P
value of
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Page 5 of 10Zhang et al. Gut Pathog (2017) 9:11
anaphylactic response and scores (Fig. 2b), and levels of
mMCP-1 (Fig. 2c), suggesting a protective effect of C.
butyricum on food allergy in mice.
Cytokines and immunoglobulinCytokines and total IgE were
determined using ELISA Kits (Fig. 3). Compared with the
control group, the lev-els of inflammatory cytokines in the serum
(Fig. 3a–d: IL-4, IL-5, IL-13, IL-17) were significantly
increased in the food allergy group and decreased in the preventive
and treated groups. However, the IL-17 of the preven-tive group was
not significant different from that of the food allergy group
(P > 0.05). Levels of INF-γ, IL-10 and TGF-β1
(Fig. 3e–g) in the food allergy group were decreased compared
with the counterparts of the con-trol group. They increased in the
groups received C. butyricum orally except for the IL-10 in the
preventive group.
Compared with the control group, the total IgE (Fig. 3h)
was significantly elevated in the food allergy group (P
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in the preventive group. Compared to the food allergy group,
Tbet expression was increased in the treated group, however, the
expression of Foxp3 remained unchanged.
Clostridium butyricum skews the immune response away
from Th2 and towards TregTo determine the extent of Th
response skewing in the spleen, ratios for Gata3/Tbet (Th2/Th1),
Foxp3/Rorγt (Treg/Th17), Foxp3/Gata3 (Treg/Th2) and Foxp3/Tbet
(Treg/Th1) mRNA expression were calculated (Fig. 6). The food
allergy groups showed a Th2-skewed immune response represented by a
significant increase in Gata3/Tbet ratio as compared to control
group. C. butyricum reduced this ratio in the treated group, but
not the preventive group. Interestingly, the Foxp3/Rorγt (except
for the treated group)and Foxp3/Gata3 ratios were significantly
increased in the groups received C. butyricum orally as compared to
the food allergy group indicating an increase in Treg-associated
responses. The ratio of Foxp3/Tbet did not differ signifi-cantly
among the different treatment groups.
The effect of C. butyricum on Treg cellWe examined the
potential induction of Foxp3 by regula-tory T cells in the MLN of
the mice. Food allergy mice had a lower percentage of CD4+
CD25+ Foxp3+ T cells in the MLN compared to
nonsensitized controls. The numbers of CD4+ CD25+
Foxp3+ Treg cells was increased in the preventive group,
however, no signifi-cant differences in these regulatory T-cell
populations were observed in the treated group as compared to the
food allergy group (Fig. 7).
Clostridium butyricum reduces histological changes
in allergic miceIngestion of BLG induced submucosal edema and
increased inflammatory cell infiltration in the gut of sensitized
mice (Fig. 8a). Allergic mice had decrease vil-lus height
whereas C. butyricum-treatment animals had a similar alteration to
the one found in the control mice (Fig. 8b). Thus, oral
administration of C. butyricum reduced the development of
intestinal inflammation.
DiscussionFood allergy is an increasing health problem and it
has a significant impact on the health and daily activities of
allergic individuals. The aim of this study was to inves-tigate the
preventive and therapeutic effect of C. butyri-cum on food allergy
in mice. In our study, there were higher scores about diarrhea and
anaphylactic symptoms in the food allergy group as compared to the
control group. Positive incidence of mice with allergic diarrhea
and anaphylactic symptoms was increased in the food allergy mice.
Additionally, the expression of mMCP-1 and total IgE were
significantly increased in the aller-gic mice. These were similar
to the results observed in a study of ovalbumin allergy [31],
however, we lack the data about BLG specific IgE due to the
unavailable of the
Fig. 5 Clostridium butyricum up-regulates Tbet and Foxp3 mRNA
expression. The mRNA expression of Th specific transcription
factors was measured by quantitative real-time PCR. Male BALB/c
mice were given NS or sensitized/challenged with BLG ± treatment
with C. butyricum. Con, n = 10 for the control group; FA, n = 10
for the food allergy group; Pre, n = 10 for the preventive group;
and Tre, n = 10 for the treated group. Results are shown as means ±
SEMs. *P < 0.05, **P < 0.01, ***P < 0.001 versus the
control group, and #P < 0.05, ##P < 0.01, ###P < 0.001
versus the food allergy group
Fig. 6 Clostridium butyricum skews the immune response away
fromTh2 and towards Treg. Ratios for Gata3/Tbet (Th2/Th1),
Foxp3/Rorγt (Treg/Th17), Foxp3/Gata3 (Treg/Th2) and Foxp3/Tbet
(Treg/Th1) mRNA expression in whole spleen tissue are shown for
each C. butyricum treatment. Male BALB/c mice were given NS or
sensitized/challenged with BLG ± treatment with C. butyricum. Con,
n = 10 for the control group; FA, n = 10 for the food allergy
group; Pre, n = 10 for the preventive group; and Tre, n = 10 for
the treated group. Results are shown as means ± SEMs. *P < 0.05,
**P < 0.01, ***P < 0.001 versus the control group, and #P
< 0.05, ##P < 0.01, ###P < 0.001 versus the food allergy
group
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Page 7 of 10Zhang et al. Gut Pathog (2017) 9:11
kit from the manufacturer. More importantly, decreased
villus length and increased inflammatory reaction were observed in
the gut slides of the food allergy mice. Hence, we successfully
mimic BLG-induced allergic intestine inflammation in our mouse
model.
Various strains of probiotics have been used in ani-mal models
of food allergy. Probiotic VSL#3-induced TGF-β ameliorates food
allergy inflammation in a mouse model of peanut sensitization
through the induction of regulatory T cells in the gut mucosa [32].
A combi-nation of specific immunotherapy with C. butyricum
significantly enforces the therapeutic effect on inhibit-ing the
food allergen related inflammation in the intes-tine, which can be
a novel approach for the treatment of food allergy [33]. A recent
study suggests that oral supplementation of Lactobacillus paracasei
L9 (L9) can reduce the development of allergic sensitization to
BLG, likely through regDCs mediated active suppression [34]. In a
BALB/c mouse model of BLG allergy, oral admin-istration of
Lactobacillus acidophilus (L. acidophilus)can suppress the major
allergic symptoms probably due to improve the regulatory T
(Treg)/Th17 balance and inhibit the IL-6 production [35]. It has
been confirmed that Bifidobacterium longum BBMN68 (BBMN68) may
be a suitable therapeutic approach to the alleviation of food
allergies likely through the specific induction of
CD11c+ CD103+ DCs and semi-mature DCs [36]. Simi-larly,
oral administration of C. butyricum to the mouse models of food
allergy is effective in reducing allergic inflammation in our
study.
Th2 cells play a key role in the pathogenesis of food allergy,
and patients with food allergy were reported to have Th1/Th2
imbalances as well as disturbed Th17/Treg balances. Th2 cytokines
including IL-4, IL-5, IL-13 [37]. IFN-γ released by Th1 can inhibit
the development of Th2. Th17 cells release IL-17 which is connected
with inflammation in the gut [38, 39]. High levels of IL-10, TGF-β
expressed by regDCs can directly mediate the conversion of T cells
into Foxp3+ Treg cells and induce immune tolerance [17]. Th2
dominance was observed in the food allergy group represented by a
significant decrease in Th1 and Treg transcription factors and high
Gata3/Tbet ratio. Hence, our model mimics the Th2-responses found
in food allergy. Importantly, C. butyri-cum shifted the immune
balance towards Th1 and Treg, with significantly increased
Foxp3/Rorγt and Foxp3/Gata ratios and a significantly decreased
Gata3/Tbet ratio. These findings are consistent with results of
those
Fig. 7 The effect of C. butyricum on Treg cell. The gating
strategy (a) and effect of oral administration of C. butyricum on
populations of CD4+ CD25+ Foxp3+ (b) cells in the MLN. Male BALB/c
mice were given NS or sensitized/challenged with BLG ± treatment
with C. butyricum. Con, n = 10 for the control group; FA, n = 10
for the food allergy group; Pre, n = 10 for the preventive group;
and Tre, n = 10 for the treated group. Results are shown as means ±
SEMs. *P < 0.05, **P < 0.01, ***P < 0.001 versus the
control group, and #P < 0.05, ##P < 0.01, ###P < 0.001
versus the food allergy group
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Page 8 of 10Zhang et al. Gut Pathog (2017) 9:11
previous studies. BBMN68 and L9 has been already shown to
significantly reduce BLG-specific hypersen-sitivity
reactions by suppressing the aberrant balance of
Th1/Th2 responses with increasing the number of CD4+
CD25+ Foxp3+ Treg cells [34, 40]. L. acido-philus
supplementation is capable of reducing allergic symptoms in a mouse
model of food allergy through reversing the imbalance of regulatory
T (Treg)/Th17. More importantly, the above observed effects of
ben-eficial bacteria on the Th responses are mirrored by the
detection of CD4+ CD25+ Foxp3+ Tregs in the MLN
of the animals. Tregs play a key role in balanc-ing immune
responses and it was demonstrated that increased expression of
Foxp3 in Tregs is directly associated with increased function of
these cells [41]. Preventive ingestion of C. butyricum increase the
pop-ulations of CD4+ CD25+ Foxp3+ cells in the MLN
and induced high levels of TGF-β and IL-10 in the serum, indicating
the powerful effect of probiotics on modu-lating the intestinal
immune response. These were sim-ilar to results observed in a study
of ovalbumin allergy, which showed that LGG only induced the number
of CD4+ CD25+ Foxp3+ Treg cells and TGF-β secretion
[42]. However, No significant differences in the per-centage of
CD4+ CD25+ Foxp3+ cells were observed
in the treated group as compared with the food allergy
group.
sIgA plays a protective role by antigen binding and exclusion
[43]. Food allergy may cause impaired epithe-lial barrier function,
including sIgA release into the gut lumen [44]. We observed
enhanced levels of sIgA in feces of the food allergy mice.
sIgA can mediate a potent anti-inflammatory
function following the interaction with SIGNR1 on DC which induces
an immune tolerance via regulatory T cell expansion [45].
Therefore, the increase of the protective secretory immunoglobulin
might be a regulatory mechanism triggered to counteract allergic
inflammation to BLG in the gut mucosa. Levels of sIgA enhanced in
allergic mice suggesting that inflamma-tion triggered this
modulatory component of immune response. The groups received C.
butyricum orally, on the other hand, had little inflammatory
consequences and therefore were not accompanied by augmented levels
of sIgA.
ConclusionsTo our knowledge, this is the first report in which
the preventive and therapeutic effects of C. butyricum on food
allergy were investigated. The findings reported here indicate that
oral probiotics such as C. butyricum, with
Fig. 8 Clostridium butyricum reduces histological changes in
allergic mice. Representative H&E staining of intestinal
sections (a, 200× magnifica-tion). Bar graph of villus height (b)
determined by measuring vertically well-oriented crypt villus units
from H&E-stainedsections of mice. Male BALB/c mice were given
NS or sensitized/challenged with BLG ± treatment with C. butyricum.
Con, n = 10 for the control group; FA, n = 10 for the food allergy
group; Pre, n = 10 for the preventive group; and Tre, n = 10 for
the treated group. Results are shown as means ± SEMs. *P < 0.05,
**P < 0.01, ***P < 0.001 versus the control group, and #P
< 0.05, ##P < 0.01, ###P < 0.001 versus the food allergy
group
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Page 9 of 10Zhang et al. Gut Pathog (2017) 9:11
established anti-inflammatory and anti-allergic activity, can
significantly modulate the mucosal immune response and may
represent an effective and safe strategy for patients with food
allergies, for which no established and effective cures based on
the pathogenetic mechanisms are available.
AbbreviationsC. butyricum: Clostridium butyricum CGMCC0313-1;
BLG: ß-lactoglobulin; Th: T helper; sIgA: secretory IgA; DC:
dendritic cells; RORγt: retinoic acid orphan receptor-γt; Foxp3:
forkhead box P3; CTX: cholera toxin; NS: normal saline; mMCP-1:
mouse mast cell protease 1; ELISA: enzyme-linked immunosorbent
assay; Ig: immunoglobulin; MLN: mesenteric lymph nodes; H&E:
hematoxylin and eosin; L9: Lactobacillus paracasei L9; regDCs:
regulatory dendritic cells; L. acidophilus: Lactobacillus
acidophilus; BBMN68: bifidobacterium longum BBMN68; SEMs: standard
error means.
Authors’ contributionsJZ, HS, YZ and XS participated in the
conception and design of the study. JZ, HS, QL, HW, ML and JH
performed laboratory work. JZ, HS, QL, HW and MZ analyzed the data
and wrote the manuscript. JZ, HS, XS and YZ contributed to the
analysis and helped in the manuscript discussion. All authors read
and approved the final manuscript.
Author details1 Department of Pediatrics, Xijing Hospital, the
Fourth Military Medical Uni-versity, Xi’an 710032, China. 2
Department of Geratology, Xijing Hospital, the Fourth Military
Medical University, Xi’an 710032, China. 3 Respiratory Depart-ment,
Shenzhen Children’s Hospital, Shenzhen 518036, China.
AcknowledgementsThe authors would like to acknowledge and thank
our funding sources.
Competing interestsThe authors declare that they have no
competing interests.
Availability of data and materialsThe datasets during and/or
analyzed during the current study are available from the
corresponding author on reasonable request.
Ethics approval and consent to participateExperimental
procedures were approved by the Ethics Committee for Animal Studies
of the Fourth Military Medical University (20150901) and performed
in accordance with their guidelines of the Institutional Animal
Care and Use Committee.
FundingThis work was supported by the innovation of science and
Technology Commission of Shenzhen Municipality
(JCYJ20120828092009036) and the National Natural Science Foundation
(31371151).
Received: 23 December 2016 Accepted: 16 February 2017
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Oral administration of Clostridium butyricum CGMCC0313-1
inhibits β-lactoglobulin-induced intestinal anaphylaxis in a
mouse model of food allergyAbstract Background: Methods:
Results: Conclusions:
BackgroundMethodsMiceThe probioticsMouse model of food
allergyMeasurement of intestinal anaphylaxisThe measurement
of cytokines and immunoglobulinSecretory IgARNA isolation
and quantitative real-time PCRFlow cytometryHistological
analysisStatistical analysis
ResultsClostridium butyricum ingestion inhibits the development
of intestinal anaphylaxisCytokines
and immunoglobulinClostridium butyricum supress the intestinal
levels of sIgAClostridium butyricum results in a strong
up-regulation of mRNA for Tbet and Foxp3Clostridium
butyricum skews the immune response away from Th2
and towards TregThe effect of C. butyricum on Treg
cellClostridium butyricum reduces histological changes
in allergic mice
DiscussionConclusionsAuthors’ contributionsReferences